卵巢癌细胞系SKOV3冻融抗原诱导CTL特异性杀瘤效应的研究  

Anti-tumor Effect and its antigenic specificity of Cytotoxicity T Lymphocytes Induced by SKOV3 Ovarian Carcinoma Cell Line Freeze-thaw Antigen

作　　者：程琪[1] 朱长焜[1] 叶枫[1] 陈怀增[1] 谢幸[1] 

机构地区：[1]浙江大学医学院附属妇产科医院,310006

出　　处：《实用癌症杂志》2005年第2期113-115,119,共4页The Practical Journal of Cancer

基　　金：浙江省科技厅重点项目基金资助项目(编号J30119)

摘　　要：目的探讨卵巢癌冻融抗原对T淋巴细胞的增殖作用及其诱导的细胞毒性T淋巴细胞(CTL)在体外杀伤卵巢癌细胞的抗原特异性细胞毒性效应。方法采用免疫磁珠分离法(magneticactivatedcellsorting,MACS)分离纯化正常人外周血CD+3T细胞,体外以SKOV3卵巢癌细胞中提取的可溶性肿瘤抗原加以刺激。溴标法检测肿瘤抗原对T细胞增殖的影响;MTT法测定肿瘤抗原诱导的CTL体外杀伤卵巢癌细胞的能力;利用绒毛膜癌细胞抗原对比观察肿瘤抗原诱导CTL杀伤肿瘤细胞的抗原特异性。结果卵巢癌细胞抗原有很强的促进T淋巴细胞增殖的作用,且存在时间依赖性,在24h时促进作用最强。SKOV3抗原诱导的CTL在体外对SKOV3细胞的杀伤率为68.38%,显著高于它对JAR绒毛膜癌细胞的杀伤率(18.31%);而JAR细胞抗原诱导的CTL对JAR和SKOV3细胞的杀伤率分别为61.02%和14.79%,有显著的抗原特异性(P=0.0001)。结论卵巢癌细胞冻融抗原诱导的CTL在体外具有很强的增殖能力和抗原特异性杀伤卵巢癌细胞的作用。Objective To investigate the proliferation,anti-tumor effect,and its antigenic specificity of CTL induced by soluble ovarian carcinoma freeze-thaw antigen.Methods CD^+_3 T cells were islolated from normal peripheral blood by high-gradient magnetic activated cell sorting(MACS) and were induced with soluble ovarian carcinoma freeze-thaw antigen generated from SKOV3 ovarian carcinoma cell line by freeze-thaw method.The proliferation of T cells stimulated by ovarian carcinoma freeze-thaw antigen was evaluated by Cell Proliferation ELISA(BrdU).In vitro cytotoxicity of CTL induced by ovanrian freeze-thaw antigen was evaluated by MTT method,and its antigenic specificity was observed by comparing with the cytotoxicity of CTL induced by choriocarcinoma cells freeze-thaw antigen.Results Ovarian carcinoma cells freeze-thaw antigen had potentiality to stimulate the proliferation of CTL in a time-dependent manner,and the peak of stimulation was reached at 24 h.CTL induced by ovarian carcinoma freeze-thaw antigen had more effective cytotoxicity against SKOV3(68.38%) than aganist JAR choriocarcinoma cells(18.31%).Meanwhile,CTL induced by choriocarcinoma freeze-thaw antigen also had much higher cytotoxicity against JAR(61.02%) than aganist SKOV3(14.79%).Conclusion CTL induced by ovarian carcinoma freeze-thaw antigen has high proliferative ability and antigenic specificity against ovarian carcinoma cells in vitro.

关 键 词：抗原诱导 CTL 卵巢癌细胞系 杀瘤效应 SKOV3细胞 细胞毒性T淋巴细胞 抗原特异性细胞 免疫磁珠分离法 CD3^+细胞 可溶性肿瘤抗原 T淋巴细胞增殖 杀伤肿瘤细胞 体外杀伤 细胞抗原 T细胞增殖 时间依赖性 JAR细胞 特异性杀伤 

分 类 号：R378.21[医药卫生—病原生物学] R730.51[医药卫生—基础医学]